MiPNet23.01 IOC126 Innsbruck AT: Difference between revisions

From Bioblast
No edit summary
No edit summary
Line 15: Line 15:


== General information ==
== General information ==
::::The Oroboros O2k-Workshop is an advanced course which covers an introduction to basic concepts on mitochondrial membrane potential: proton pumps, ion gradients, electrochemical potential (proton motive force) and the measurement of mt-membrane potential in biological samples (isolated mitochondria, homogenate) with potentiometric (TPP+/O2k-TPP+ ISE-Module) and amperometric method (TMRM, safranin/O2k-Fluo LED2-Module) Furthermore, data analysis - from TPP+ to mt-membrane potential - constitutes a major topic. Β 
::::The Oroboros O2k-Workshop is an advanced course which covers an introduction to basic concepts on mitochondrial membrane potential: proton pumps, ion gradients, electrochemical potential (proton motive force) and the measurement of mt-membrane potential in biological samples (isolated mitochondria, homogenate) with potentiometric (O2k-TPP+ ISE-Module) and fluorescence method (TMRM, safranin/O2k-Fluo LED2-Module). Data analysis - from TPP+ to mt-membrane potential - constitutes a major topic. Β 
[[File:IOC126.png|center|500px|data analysis]]
[[File:IOC126.png|center|500px|data analysis]]


Line 49: Line 49:
|-
|-
|11:00
|11:00
|Determination of mitochondrial membrane potential (mtMP) with potentiometric TPP+/O2k-TPP+ ISE-Module - Introduction
|Determination of mitochondrial membrane potential (mtMP) with potentiometric O2k-TPP+ ISE-Module - an introduction.
|MiP''Art''
|MiP''Art''
|-
|-
Line 57: Line 57:
|-
|-
|13:15
|13:15
|Hands-on: Assembly and maintenance of TPP and reference electrode [[MiPNet15.03 O2k-MultiSensor-ISE]]
|Hands-on: Assembly and maintenance of TPP+ and reference electrodes [[MiPNet15.03 O2k-MultiSensor-ISE]]
|MiP''Art''
|MitoFit ORO Lab
|-
|-
|14:30
|14:30
|Hands-on: Set up of the instruments with TPP and reference electrodes + TPP calibration [[MiPNet15.03 O2k-MultiSensor-ISE]]
|Hands-on: Set up of the instruments with TPP+ and reference electrodes, TPP+ calibration [[MiPNet15.03 O2k-MultiSensor-ISE]]
|MitoFit ORO Lab Β 
|MitoFit ORO Lab Β 
|-
|-
Line 69: Line 69:
|-
|-
|17:00
|17:00
|Hands-on continued: TPP chemical background [[MiPNet14.05 TPP-mtMembranePotential]] Β 
|Hands-on continued: TPP+ chemical background [[MiPNet14.05 TPP-mtMembranePotential]] Β 
|MiP''Art''
|MitoFit ORO Lab
|}
|}


Line 80: Line 80:
|-
|-
|09:00
|09:00
|Introduction in planning and performing a TPP experiment; instrumental background oxygen flux in the presence of the TPP electrode O2k-background [[MiPNet14.06 Instrumental O2 background]]
|Introduction in planning and performing a TPP+ experiment.
|MiP''Art''
|MitoFit ORO Lab
|-
|-
|09:30
|09:30
|Hands-on: Calibration of [[MiPNet15.08 TPP electrode]]
|Hands-on: Calibration of TPP+ electrode.
|MitoFit ORO Lab Β 
|MitoFit ORO Lab Β 
|-
|-
Line 92: Line 92:
|-
|-
|11:00
|11:00
|Hands-on continued TPP: experiment with mouse brain mitochondria
|Hands-on continued TPP+: experiment with mouse brain mitochondria.
|MitoFit ORO Lab Β 
|MitoFit ORO Lab Β 
|-
|-
Line 101: Line 101:
|14:00
|14:00
| From the TPP signal to mt-membrane potential - Guide through the Excel templates. [[Media:MiPNet14.05_App_TPP-Mathematics-MembranePotential.pdf|TPP-mtMembranePotential]] mathematics
| From the TPP signal to mt-membrane potential - Guide through the Excel templates. [[Media:MiPNet14.05_App_TPP-Mathematics-MembranePotential.pdf|TPP-mtMembranePotential]] mathematics
|MiP''Art''
|MitoFit ORO Lab
|-
|-
|14:30
|14:30
| Analysis of TPP experiment - discussion on mitochondrial membrane potential,[[Media:MiPNet14.05_App_TPP-Mathematics-MembranePotential.pdf|TPP-mtMembranePotential]] mathematics
| Analysis of TPP experiment and discussion on mitochondrial membrane potential.
|MiP''Art''
|MitoFit ORO Lab
|-
|-
|16:00
|16:00
Line 116: Line 116:
|-
|-
|17:00
|17:00
|Determination of mitochondrial membrane potential (mtMP) with amperometric method (TMRM, safranin/O2k-Fluo LED2-Module) - introduction
|Determination of mitochondrial membrane potential (mtMP) with fluorescence method (TMRM, safranin/O2k-Fluo LED2-Module) - an introduction.
|MitoFit ORO Lab Β 
|MitoFit ORO Lab Β 
|-
|-
|17:30
|17:30
|Hands-on: Set up of the instruments with LED2-Module, [[MiPNet17.05 O2k-Fluo LED2-Module]], Β 
|Hands-on: Set up of the instruments with LED2-Module, choosing Fluorescence Sensors [[MiPNet17.05 O2k-Fluo LED2-Module]], [http://wiki.oroboros.at/index.php/Fluorescence-Sensor]
Calibration of Safranin/TMRM, chemical background experiment
Calibration of Safranin/TMRM, chemical background experiment
|MitoFit ORO Lab Β 
|MitoFit ORO Lab Β 
Line 134: Line 134:
|09:00
|09:00
|Introduction in planning and performing an experiment with Safranin/TMRM Β 
|Introduction in planning and performing an experiment with Safranin/TMRM Β 
|MiP''Art''
|MitoFit ORO Lab
|-
|-
|09:30
|10:00
|Hands-on: O2k setup for air calibration
|Hands-on: O2k setup for air calibration
|MitoFit ORO Lab Β 
|MitoFit ORO Lab Β 
Line 154: Line 154:
|14:00
|14:00
|Analysis of TMRM/Safranin experiment - discussion Β 
|Analysis of TMRM/Safranin experiment - discussion Β 
|MiP''Art''
|MitoFit ORO Lab
|-
|-
|15:00
|15:30
|''Coffee/Tea''
|''Coffee/Tea''
|
|
|-
|-
|16:30
|16:00
|Round table: questions
|Comparison of TPP+ and fluorescence methods for evaluation of mitochondrial membrane potential.
|MiP''Art''
|MiP''Art''
|-
|-
|17:00
|17:00
|Other potentiometric, amperometric, and fluorometric O2k-MultiSensor applications, the Bioblast wiki, O2k-Network and MitoEAGLE, feedback and conclusions
|Other potentiometric, amperometric, and fluorometric O2k-MultiSensor applicationsΒ  Β 
|the Bioblast wiki, O2k-Network and MitoEAGLE, feedback and conclusions
|MiP''Art''
|MiP''Art''
|-
|-

Revision as of 14:17, 20 December 2017

                



MiPNet23.01 IOC126 Innsbruck AT

Publications in the MiPMap
OROBOROS-solo.png

Innsbruck AT, 2018 Jan 24-26. Oroboros advanced O2k-Workshop on mt-membrane potential: TPP+ and fluorescent dyes. IOC126.


Oroboros (2018-01-24) Mitochondr Physiol Network

Abstract:

O2k-Workshops.png

Oroboros advanced O2k-Workshop on mt-membrane potential: TPP+ and fluorescent dyes. Innsbruck, Austria; 2018 Jan 24-26.


β€’ O2k-Network Lab: AT_Innsbruck_Oroboros, AT_Innsbruck_MitoFit


General information

The Oroboros O2k-Workshop is an advanced course which covers an introduction to basic concepts on mitochondrial membrane potential: proton pumps, ion gradients, electrochemical potential (proton motive force) and the measurement of mt-membrane potential in biological samples (isolated mitochondria, homogenate) with potentiometric (O2k-TPP+ ISE-Module) and fluorescence method (TMRM, safranin/O2k-Fluo LED2-Module). Data analysis - from TPP+ to mt-membrane potential - constitutes a major topic.
data analysis


Venue

Information on travel and venue:IOC Innsbruck.
Β» Hotel suggestion: Hotel Weisses Kreuz
Mention "Oroboros" when booking in order to receive a discount.

Preliminary programme

Wednesday 24th Topic Room
09:00 Get-together: Introduction of participants and their research interests MiPArt
09:30 Erich Gnaiger: Protonmotive force and chemiosmotic pressure: a generalization of non-ohmic flux control of the proton leak. See: Abstract. MiPArt
10:00 O2k-MultiSensor overview, DatLab 7, DatLab 7 Protocols MiPArt
10:30 Coffee/Tea
11:00 Determination of mitochondrial membrane potential (mtMP) with potentiometric O2k-TPP+ ISE-Module - an introduction. MiPArt
11:45 Lunch break
13:15 Hands-on: Assembly and maintenance of TPP+ and reference electrodes MiPNet15.03 O2k-MultiSensor-ISE MitoFit ORO Lab
14:30 Hands-on: Set up of the instruments with TPP+ and reference electrodes, TPP+ calibration MiPNet15.03 O2k-MultiSensor-ISE MitoFit ORO Lab
16:30 Coffee/Tea
17:00 Hands-on continued: TPP+ chemical background MiPNet14.05 TPP-mtMembranePotential MitoFit ORO Lab
Thursday 25th Topic Room
09:00 Introduction in planning and performing a TPP+ experiment. MitoFit ORO Lab
09:30 Hands-on: Calibration of TPP+ electrode. MitoFit ORO Lab
10:30 Coffee/Tea
11:00 Hands-on continued TPP+: experiment with mouse brain mitochondria. MitoFit ORO Lab
12:30 Lunch break
14:00 From the TPP signal to mt-membrane potential - Guide through the Excel templates. TPP-mtMembranePotential mathematics MitoFit ORO Lab
14:30 Analysis of TPP experiment and discussion on mitochondrial membrane potential. MitoFit ORO Lab
16:00 Coffee/Tea
16:30 Hands-on: O2k setup for air calibration MitoFit ORO Lab
17:00 Determination of mitochondrial membrane potential (mtMP) with fluorescence method (TMRM, safranin/O2k-Fluo LED2-Module) - an introduction. MitoFit ORO Lab
17:30 Hands-on: Set up of the instruments with LED2-Module, choosing Fluorescence Sensors MiPNet17.05 O2k-Fluo LED2-Module, [1]

Calibration of Safranin/TMRM, chemical background experiment

MitoFit ORO Lab
Friday 26th Topic Room
09:00 Introduction in planning and performing an experiment with Safranin/TMRM MitoFit ORO Lab
10:00 Hands-on: O2k setup for air calibration MitoFit ORO Lab
10:30 Coffee/Tea
11:00 Hands-on: Calibration of Safranin/TMRM, experiment with mouse brain mitochondria/HEK cells MitoFit ORO Lab
12:30 Lunch break
14:00 Analysis of TMRM/Safranin experiment - discussion MitoFit ORO Lab
15:30 Coffee/Tea
16:00 Comparison of TPP+ and fluorescence methods for evaluation of mitochondrial membrane potential. MiPArt
17:00 Other potentiometric, amperometric, and fluorometric O2k-MultiSensor applications the Bioblast wiki, O2k-Network and MitoEAGLE, feedback and conclusions MiPArt
18:00 Farewell activity



Registration

Registration form

Tutors and Lecturers

Tutors

Lecturers and assistance

List of participants

Preliminary list
Participant Institution
Kandel Sunil Mani US WI Milwaukee Dash RK - Medical College of Wisconsin
Smirnova Irina SE Stockholm Smirnova I - Stockholm University
Magdalena Labieniec-Watala Labieniec-Watala Magdalena PL Lodz Watala C - University of Lodz
Janssen Connie NL Leiden Lindeman JH - Leiden University Medical Center
de Kok MichΓ©le NL Leiden Lindeman JH - Leiden University Medical Center
Steffen Anja NL Leiden Lindeman JH - Leiden University Medical Center
Siewiera Karolina Siewiera Karolina PL Lodz Watala C - University of Lodz
Michalak Slawomir Michalak Slawomir PL Poznan Michalak S - Poznan University of Medical Sciences
Fernandez-Vizarra Erika UK Cambridge Zeviani M - University of Cambridge


Contact

[email protected]
Oroboros Instruments
High-Resolution FluoRespirometry
SchΓΆpfstrasse 18
A-6020 Innsbruck, Austria
Tel: +43 512 566796
Fax: +43 512 566796 20
Mitochondria and cell research

IOC recommended reading

Β» IOC recommended reading
Β» MitoPedia: Respiratory states
OXPHOS ROUTINE ETS LEAK ROX

Project support

Contribution to K-Regio project MitoFit, funded by the Land Tirol within the program K-Regio of Standortagentur Tirol.
Sponsors.png


Labels:






ORO, IOC, MitoFit, Next, 2018 


MitoGlobal
O2k-Workshops and MitoFit Coaching Days are listed as MitoGlobal Events.
Cookies help us deliver our services. By using our services, you agree to our use of cookies.